hCD73(2)

Fig.1 Detection of NT5E expression in liver and lung by RT-PCR. 

Wild type: only one band at 221 bp with primers F1/R1(mNt5e);

Homozygous: only one band at 285 bp with primers F2/R2(hNT5E); 

Abbr.. M, DNA marker; HO, homozygous; HE, heterozygous; WT, wild type.

Fig.2 Detection of CD73 expression on T cells in spleen of WT C57BL/6 mice and HO hCD73(2) mice by FACS. Leukocytes were collected from wild-type C57BL/6 mice (female, 3 months old) and homozygous hCD73(2) humanized mice (male, 12 weeks old), and analyzed by flow cytometry with anti-mCD73 and anti-hCD73. hCD73 was exclusively detectable in hCD73(2) humanized mice but not in wild-type mice. hCD73 was exclusively detectable on T cells in hCD73(2) humanized mice, whereas mCD73 was exclusively detectable on T cells in wild-type C57BL/6 mice.

Abbr. HO, homozygous; WT, wild type.

Fig.3 Detection of CD73 expression on T cells in bone marrow of WT C57BL/6 mice and HO hCD73(2) mice by FACS. Leukocytes were collected from wild-type C57BL/6 mice (female, 3 months old) and homozygous hCD73(2) humanized mice (male, 12 weeks old), and analyzed by flow cytometry with anti-mCD73 and anti-hCD73. hCD73 was exclusively detectable in hCD73(2) humanized mice but not in wild-type mice. hCD73 was exclusively detectable on T cells in hCD73(2) humanized mice, whereas mCD73 was exclusively detectable on T cells in wild-type C57BL/6 mice.

Abbr. HO, homozygous; WT, wild type.

Fig.4 Expression of CD73 in the  peripheral T cells collected from humanized CD73 mice were detected by FACS. The results showed that the expression of human CD73 can be detected in the  peripheral T cells of humanized CD73 mice as same as in Hu-PBMC.

Fig.5 In vivo validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized CD73 mice. Homozygous humanized CD73 mice were inoculated with MC38 colon cancer cells (expressing human CD73 rather than murine CD73). Mice were grouped when the tumor size was approximately 100 mm3(n=6). Three human CD73 antibodies differently inhibited tumor growth, confirming that the CD73-HU mouse model is a powerful tool for in vivo CD73 antibody pharmacological efficacy study. A.Tumor average volume ± SEM, B.Mice average weight ± SEM.